Effect of ADM siRNA on the expression ofβ-catenin and terminal differentiation of osteosarcoma cells / 西安交通大学学报(医学版)

ABSTRACT

Objective To observe the effect of silencing the expression of ADM using RNA interfering technique on the expression ofβ-catenin and terminal differentiation of osteosarcoma cells.Methods After the intervention of 0 h,48 h and 72 h by ADM siRNA,we observed the change of distribution ofβ-catenin in F5M2 using immunocytochemistry staining.The expression levels of total and phosphorylatedβ-catenin and GSK3βwere detected by Western blot after the intervention by ADM siRNA.The F5 M2 cells treated with ADM siRNA were subjected to HE staining,alkaline phosphatase (ALP)assay and immunocytochemistry staining to investigate the biological effects of ADM siRNA on the morphology and terminal differentiation of F5M2 cells.Results After the intervention of 0 h,48 h and 72 h by ADM siRNA,the distribution ofβ-catenin was transferred from the cytoplasm to the nucleus.ADM siRNA downregulated the expression level of P-β-catenin and upregulated P-GSK3βdetected by Western blot.HE staining revealed that the configuration of F5M2 had undergone restorational changes similar to those of normal cells after ADM siRNA treatment.ALP assay and immunocytochemistry staining showed that the expression of the earlier and later molecular biomarkers of terminal differentiation,including ALP and osteocalcin were strongly positive.Conclusion Silencing the expression of ADM can activateβ-catenin to transfer to the nucleus from the cytoplasm,and induce osteosarcoma cells to make terminal differentiation through activatingβ-catenin signaling pathway.